Objective The effects of carrimycin(CAM)on the biological functions of intrahepatic cholan-giocarcinoma HuCCT1 cells were examined through in vitro experiments,and a preliminary investigation was conducted into its mechanism of action.Methods The intrahepatic cholangiocarcinoma cell line HuCCT1 was selected for the study.The effect of CAM on cell viability was assessed using the CCK-8 assay,and the IC50 concen-tration was determined accordingly.The impact of CAM on cell migration was evaluated through a scratch wound healing assay.In addition,the effect of CAM on clonogenic ability was examined using a colony formation assay.Cell invasion capacity was assessed using a Transwell invasion assay.Flow cytometry was employed to analyze the effect of CAM on cell cycle progression.Furthermore,Western blotting was conducted to evaluate the expression levels of key proteins associated with epithelial-mesenchymal transition and the cell cycle.Finally,the influence of CAM on the AXL/c-Myc/c-Met signaling axis was also investigated.Results Compared with the control group,CAM significantly inhibited the proliferation of HuCCT1 cells in a concentration-dependent manner(P<0.05).Plate cloning assays demonstrated that CAM markedly suppressed the colony-forming ability of HuCCT1 cells(P<0.05).Scratch wound healing assays confirmed that CAM treatment significantly reduced the migration speed and narrowed the migration area of HuCCT1 cells(P<0.05).Flow cytometry analysis revealed that CAM treatment led to a significant increase in the proportion of cells in the G0/G1 phase and a decrease in the S phase(P<0.05).Western blot analysis further confirmed that the expression levels of key regulatory proteins CCND1 and CDK4,which are involved in the G1/S phase transition,were down-regulated,while the expression of p21 was up-regulated(P<0.05).Transwell invasion assays indicated that CAM inhibited the invasive capacity of HuCCT1 cells.Consis-tently,Western blot results showed that E-Cadherin expression was increased(P<0.05),whereas the expression levels of N-Cadherin and Vimentin were decreased(P<0.05).Moreover,Western blot analysis verified that the expression of AXL,c-Met,and c-Myc was up-regulated in HuCCT1 cells treated with AXL recombinant protein(P<0.05).However,co-treatment with CAM and AXL recombinant protein significantly attenuated the expression of these proteins(P<0.05).Conclusions CAM inhibits the proliferation,migration,and invasion of intrahe-patic cholangiocarcinoma HuCCT1 cells,thereby demonstrating antitumor effects,which may be associated with the AXL/c-Met/c-Myc signaling pathway.

Objective The effects of carrimycin(CAM)on the biological functions of intrahepatic cholan-giocarcinoma HuCCT1 cells were examined through in vitro experiments,and a preliminary investigation was conducted into its mechanism of action.Methods The intrahepatic cholangiocarcinoma cell line HuCCT1 was selected for the study.The effect of CAM on cell viability was assessed using the CCK-8 assay,and the IC50 concen-tration was determined accordingly.The impact of CAM on cell migration was evaluated through a scratch wound healing assay.In addition,the effect of CAM on clonogenic ability was examined using a colony formation assay.Cell invasion capacity was assessed using a Transwell invasion assay.Flow cytometry was employed to analyze the effect of CAM on cell cycle progression.Furthermore,Western blotting was conducted to evaluate the expression levels of key proteins associated with epithelial-mesenchymal transition and the cell cycle.Finally,the influence of CAM on the AXL/c-Myc/c-Met signaling axis was also investigated.Results Compared with the control group,CAM significantly inhibited the proliferation of HuCCT1 cells in a concentration-dependent manner(P<0.05).Plate cloning assays demonstrated that CAM markedly suppressed the colony-forming ability of HuCCT1 cells(P<0.05).Scratch wound healing assays confirmed that CAM treatment significantly reduced the migration speed and narrowed the migration area of HuCCT1 cells(P<0.05).Flow cytometry analysis revealed that CAM treatment led to a significant increase in the proportion of cells in the G0/G1 phase and a decrease in the S phase(P<0.05).Western blot analysis further confirmed that the expression levels of key regulatory proteins CCND1 and CDK4,which are involved in the G1/S phase transition,were down-regulated,while the expression of p21 was up-regulated(P<0.05).Transwell invasion assays indicated that CAM inhibited the invasive capacity of HuCCT1 cells.Consis-tently,Western blot results showed that E-Cadherin expression was increased(P<0.05),whereas the expression levels of N-Cadherin and Vimentin were decreased(P<0.05).Moreover,Western blot analysis verified that the expression of AXL,c-Met,and c-Myc was up-regulated in HuCCT1 cells treated with AXL recombinant protein(P<0.05).However,co-treatment with CAM and AXL recombinant protein significantly attenuated the expression of these proteins(P<0.05).Conclusions CAM inhibits the proliferation,migration,and invasion of intrahe-patic cholangiocarcinoma HuCCT1 cells,thereby demonstrating antitumor effects,which may be associated with the AXL/c-Met/c-Myc signaling pathway.

Amyotrophic Lateral Sclerosis (ALS) is a complex neurodegenerative disorder characterized by progressive axonopathy, jointly leading to the dying back of the motor neuron, disrupting both nerve signaling and motor control. In this review, we highlight the roles of axonopathy in ALS progression, driven by the interplay of multiple factors including defective trafficking machinery, protein aggregation, and mitochondrial dysfunction. Dysfunctional intracellular transport, caused by disruptions in microtubules, molecular motors, and adaptors, has been identified as a key contributor to disease progression. Aberrant protein aggregation involving TDP-43, FUS, SOD1, and dipeptide repeat proteins further amplifies neuronal toxicity. Mitochondrial defects lead to ATP depletion, oxidative stress, and Ca²⁺ imbalance, which are regarded as key factors underlying the loss of neuromuscular junctions and axonopathy. Mitigating these defects through interventions including neurotrophic treatments offers therapeutic potential. Collaborative research efforts aim to unravel ALS complexities, opening avenues for holistic interventions that target diverse pathological mechanisms.
Humans ; Amyotrophic Lateral Sclerosis/therapy* ; Animals ; Axons/metabolism* ; Mitochondria/metabolism* ; Motor Neurons/pathology*

Objective The aim of this study was to investigate the expression of miR-455-5p in epithelial ovarian cancer and its effect on the development of epithelial ovarian cancer. Methods The miRNA expression data of normal ovarian epithelial tis-sues and epithelial ovarian cancer tissues GSE83693 were downloaded from the GEO database. Differential expression analysis was used to obtain differential expression data of miRNAs in epithelial ovarian cancer. The expression of miR-455 -5p was analyzed whether there is difference expression between normal ovarian epithelium and epithelial ovary cancer tissues; qRT-PCR was used to verify the differential expression prediction results; bio-informatics software was used to analyze the KEGG pathway enrichment and GO gene function annotation of miR-455-5p target genes,and to explore the disorders of dyregulated miR-455-5p in the devel-opment of epithelial ovarian cancer. Results A total of 101 cases of differentially expressed miRNAs were screened,34 cases were up-regulated and 67 cases were down-regulated. Among them,miR-455-5p was down-regulated significantly(P<0. 01),and the different fulds were -2. 9019. The results of qRT-PCR showed that the expression of miR-455-5p in epithelial ovarian cancer cells(SKOV-3,OVCAR-3 and A2780)was significantly lower than that in normal ovarian epithelial cells(IOSE-80),and the dif-ferential expression was statistically significant(P<0. 05). The results of KEGG pathway enrichment analysis showed that miR-455-5p regulated target genes mainly involved in five pathways,including TGF-β signaling pathway,Hippo signaling pathway,ECM-receptor interaction,transcriptional dysregulation pathway in cancer,and chronic granule cellular leukemia,which were associated with tumors. GO functional annotation analysis showed that the target genes regulated by miR-455-5p in the above pathway was mainly involved in protein phosphorylation,promoted cell proliferation and migration,inhibited apoptosis,promoted epithelial-mesenchymal transition,regulated transcription and regulated cell cycle,etc. ,which associated with tumorigenesis. Conclusion The expression of miR-455-5p is down-regulated in epithelial ovarian cancer. The miR-455-5p target genes are involved in the pathogenesis and function of epithelial ovarian cancer,and are associated with the development of epithelial ovarian cancer.

Objective The aim of this study was to investigate the expression of microRNA-150(miR-150)in human epi-thelial ovarian cancer cells and its effect on proliferation,apoptosis,invasion and metastasis of human epithelial ovarian cancer cells. Methods The expression level of miR-150 in cells from each treatment group was detected by Real-Time PCR(qRT-PCR);effects of proliferation,apoptosis,invasion and metastasis of epithelial ovarian cancer cells was investigated by MTT,flow cytometry, and transwell assays. Results Compared with normal ovarian epithelial cells(T29),the expression of miR-150 was significantly de-creased in epithelial ovarian cancer cells(A2780 and OVCAR3)(P<0. 01); After transfection miR-150 mimic,the expression of miR-150 in A2780 and OVCAR3 cells was significantly increased(P<0. 01);After 3 d of transfection,the OD values of the miR-150mimicgroup(A2780:1.12±0.03;OVCAR3:1.91±0.03)werelowerthanthatintheblankgroup(A2780:2.35±0.09;OVCAR3:2.63 ±0.07)and the miR-150 NC group(A2780:2.18 ±0.07;OVCAR3:2.43 ±0.11)(P<0.01);The apoptotic rate in the miR-150 mimic group(A2780:16. 10 ± 0. 58% ;OVCAR3:15. 16 ± 1. 30% ) were significantly increased when compared to the blank group(A2780:10. 07 ± 0. 66%;OVCAR3:3. 81 ± 0. 24%) and the miR -150 NC group(A2780:10. 36 ± 1. 08%;OVCAR3:4.89 ±0.07%)(P<0.01);The number of transmembrane cells in the miR-150 mimic group(A2780:38.67 ±2.03;OVCAR3:28. 67 ± 2. 03)was higher than that in the blank group(A2780:76. 30 ± 7. 45;OVCAR3:55. 67 ± 3. 18)and the miR-150 NC group(A2780:74. 33 ± 5. 78;OVCAR3:56. 33 ± 3. 84)(P<0. 01). Conclusion The decreased expression of miR-150 in epi-thelial cancer cells may be one of the mechanisms of proliferation,invasion and metastasis of epithelial ovarian cancer. Up-regulation of miR-150 may inhibit the proliferation of epithelial ovarian cancer cells and promote apoptosis to reduce the abilities of invasion and metastasis in epithelial ovarian cancer cells.

AIM: To observe the difference of cerebral injury following ischemia/reperfusion and mRNA expression of TGF-β1 between diabetic and non-diabetic rats.METHODS: At first, Wistar rats were divided into two groups,non-diabetes and diabetes, and then two groups followed by sham, middle cerebral artery occlusion(MCAO) 2h and reperfusion 24 h after MCAO 2 h respectively. TGF-β1 mRNA expression was measured by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR); Cerebral damage was evaluated by histopathology.RESULTS: In the same condition of ischemia or ischemia/reperfusion, injuried area enlarged in DM groups; The expression level of TGF-β1 mRNA increased at the time of 2 h after MCAO in non-diabetic group and diabetic group, especialy significantly in non-diabetic group with MCAO 2 h, and decreased at the time of reperfusion 24 h after MCAO 2 h, but still higher than that in the sham group.CONCLUSION: Diabetes mellitus exacerbated brain lesion following ischemia/repefusion, increased TGF-β1 mRNA expresion after MCAO may be an anti-injury reaction,and anti-injury ability is decreased under diabetic condition

AIM: To observe the difference of cerebral injury following ischemia/reperfusion and mRNA expression of TGF-? 1 between diabetic and non-diabetic rats.METHODS: At first, Wistar rats were divided into two groups,non-diabetes and diabetes, and then two groups followed by sham, middle cerebral artery occlusion(MCAO) 2h and reperfusion 24 h after MCAO 2 h respectively. TGF-? 1 mRNA expression was measured by semi-quantitative reverse transcription polymerase chain reaction(RT-PCR); Cerebral damage was evaluated by histopathology.RESULTS: In the same condition of ischemia or ischemia/reperfusion, injuried area enlarged in DM groups; The expression level of TGF-? 1 mRNA increased at the time of 2 h after MCAO in non-diabetic group and diabetic group, especialy significantly in non-diabetic group with MCAO 2 h, and decreased at the time of reperfusion 24 h after MCAO 2 h, but still higher than that in the sham group.CONCLUSION: Diabetes mellitus exacerbated brain lesion following ischemia/repefusion, increased TGF-? 1 mRNA expresion after MCAO may be an anti-injury reaction,and anti-injury ability is decreased under diabetic condition.

AIM: To study forms of cell death following cerebral ischemia/reperfusion in diabetic rats. METHODS: Based on the modles of diabetes and middle cerebral artery occlusion(MCAO), characteristics of cell death after ischemia/reperfusion were evaluated synthetically by the pathological, flow cytometry(FCM), TUNEL and the DNA agarose electrophoresis.RESULTS: The occurrence of cerebral injury after ischemia/reperfusion were accompanied by cell necrosis and cell apoptosis. And cell apoptosis was mainly located in ischeamic penumbra(IP) zone around the densely ischemic focus. Ischemic centre(IC)was characterized by cell necrosis. At the same time, the results showed that the process of ischemic cerebral injury worsen by diabetes mellitus was related to inducing cell apoptosis in IP and Mid zone.CONCLUSION: Neuronal damage following focal cerebral ischemia/reperfusion included cell necrosis and apoptosis, IC zone was mainly characterized by the former, however IP zone by the latter, and there had close internal relationship between them. Brain damage following cerebral ischemia/reperfusion was worsen instinctly under diabetic condition.